吡嗪酰胺耐药的耐多药结核分枝杆菌pncA基因表达分析  

Analysis of pncA gene expression in pyrazinamide-resistant and multidrug-resistant Mycobacterium tuberculosis

作　　者：王楠[1] 杨瑜[1] 邓丽 吴玲[1] 刘志辉[1] 孟繁荣[1] Wang Nan;Yang Yu;Deng Li;Wu Ling;Liu Zhihui;Meng Fanrong(.Institue of Pulomonary Diseases,Guangzhou Chest Hospital,State Key Laboratory of Respiratory Disease,Guangzhou 510095,China)

机构地区：[1]广州市胸科医院肺部疾病研究所呼吸疾病国家重点实验室,广州510095

出　　处：《新医学》2023年第12期879-883,共5页Journal of New Medicine

基　　金：广东省医学科学技术研究基金项目(A2023284);广州市科技计划项目(202201010697);广州市卫生健康科技项目(20241A011049);广州市结核病研究重点实验室(2023A03J0539)。

摘　　要：目的 比较吡嗪酰胺(PZA)耐药的耐多药结核分枝杆菌在PZA药物刺激前后PZA耐药基因pncA的表达,探讨pncA基因表达与结核分枝杆菌PZA耐药的关系。方法 培养PZA耐药的耐多药结核分枝杆菌,检测其利福平和PZA最小抑菌浓度(MIC);以PZA浓度为0和50μg/mL的培养基培养菌株,实时荧光定量PCR检测菌株pncA的表达水平。结果 21株结核分枝杆菌PZA的MIC≥1 600μg/mL、12株为400~800μg/mL、5株为100~200μg/mL;分别定义相应PZA耐药水平为高、中、低,其利福平的MIC中位数分别为128、128、64μg/mL,任意2组利福平的MIC比较差异均无统计学意义(P均> 0.05);以相对定量2^(-ΔΔCt)法计算PZA高、中、低水平耐药结核分枝杆菌pncA表达水平,无药组分别为0.904(0.202,2.653)、1.157(0.658,1.511)、1.147(0.372,1.347),PZA浓度为50μg/mL处理组分别为1.544(0.611,3.191)、0.846(0.421,1.237)、0.726(0.225,3.017);以无药组为对照,PZA含药培养组与其对比,pncA基因表达差异无统计学意义(P> 0.05)。结论 PZA耐药的耐多药结核分枝杆菌在PZA药物刺激前后pncA表达无明显变化。Objective To compare the expression levels of pncA gene in pyrazinamide(PZA)-resistant and multidrug-resistant Mycobacterium tuberculosis before and after PZA drug stimulation,and to explore the relationship between PZA-resistant Mycobacterium tuberculosis and pncA gene expression.Methods PZA-resistant and multidrug-resistant Mycobacterium tuberculosis was cultured,and the minimal inhibitory concentration(MIC)of rifampicin and PZA was detected.The strains were cultured in medium containing 0 and 50μg/mL PZA,and the expression level of pncA was detected by real-timefluorescence quantitative PCR.Results The MIC of PZA of 21 strains of Mycobacterium tuberculosis was≥1600μg/mL,400-800μg/mL for 12 strains,and 100-200μg/mL in 5 strains,and their corresponding PZA resistance levels were defined as high,medium and low,respectively.The median MIC of rifampicin was 128,128 and 64μg/mL,respectively.Chi-square test showed that there was no significant difference between any two groups(all P>0.05).According to the relative quantitative 2-ΔΔCt method,the expression levels of pncA in the high,medium and low levels of PZA drug-resistant Mycobacterium tuberculosis was 0.904(0.202,2.653),1.157(0.658,1.511)and 1.147(0.372,1.347),and 1.544(0.611,3.191),0.846(0.421,1.237)and 0.726(0.225,3.017)in the 50μg/mL treatment group,respectively.There was no significant difference in pncA gene expression between the control and PZA drug-containing culture groups(P>0.05).Conclusion No significant changes are observed in the expression levels of pncA in multidrug-resistant and PZA-resistant Mycobacterium tuberculosis before and after PZA drug stimulation.

关 键 词：结核分枝杆菌 耐药 吡嗪酰胺 利福平 PNCA 

分 类 号：R378.911[医药卫生—病原生物学]